Improved Taxol Yield in Cell Suspension Culture ofTaxus wallichiana(Himalayan Yew)

Abstract

Cell culture of Taxus wallichiana Zucc (= T. baccata ssp wallichiana Zucc. Pilg.) (Himalayan yew) has been established and taxol-producing cell lines selected by cell line cloning. Cell line NC110 derived from needle leaf of a 40-year-old tree growing in Darjeeling Himalayas produced 0.018% taxol in B5 basal medium supplemented with 2,4-D (2 mg/l), kinetin (0.5 mg/l), and casein hydrolysate (0.5%). This cell line has been maintained for over 2 years. Significant enhancement in the level of taxol (0.05%) was obtained in this cell line by supplementation of the basal medium with 5 mg/l of IAA-phenylalanine instead of 2,4-D without adversely affecting cell growth. IAA-glycine also enhanced taxol level (0.03%) while IAA alone (1-10 mg/l) was ineffective in inducing taxol accumulation. Using three different cell lines with different taxol-producing capacities, it has been demonstrated that 2,4-D and IAA-phenylalanine when present alone favour growth and taxol production but when combined enhance biomass to a maximum (six-fold in NC110) without enhancing taxol accumulation, suggesting that a two-stage culture may be beneficial for optimising taxol accumulation in cell culture of T. wallichiana.

Detection of anthraquinones and identification of 1,4-naphthohydroquinone in cell suspension cultures of Rudgea jasminoides (Cham.) Müll. Arg. (Rubiaceae)

“…A number of plant species have been used for generation and propagation of cell-suspension cultures, ranging from model systems like Taxus (Jha et al 1998), Arabidopsis (Richard et al 2001) and Catharanthus (Kim et al 2004) to important monocotyledon or dicotyledon crop plants like soybean (Chu & Lark 1976), alfalfa (Atanassov & Brown 1984), rice (Lee et al 2004), and tobacco (Hellwig et al 2004).…”

Recent advances in plant biotechnology and genetic engineering for production of secondary metabolites

“…Results of these studies were sum marized in recent publications [43,44]. Manipu lation with cultural medium composition in com bination with efficient selection allowed in a num ber of cases accumulation of paclitaxel in cells up to 0.03-0.05 % d. w. that is comparable or even surpasses the metabolite level in T. brevifolia bark [45,46]. Further investigation proved efficiency of elicitation for taxoid biosynthesis stimulation because a number of important enzymes of ter pene pathway (for instance geranylgeranyl diphos phate synthase and taxadiene synthase) are jas monate inducible [47,48].…”

Secondary metabolites production through biotechnological intervention: A Review

“…The first time, in vitro taxol production was carried out by the Christen et al (1989), thereafter, similar approached has been adopted in severeral research laboratories throughtout the world (Flores and Sgrignoli, 1991;Ma et al, 1994;Lee et al, 1995;Yukimune et al, 1996;Jha et al 1998;Nguyen et al, 2001;Wu et al, 2001;Linden et al, 2001;Parc et al, 2002;Abbasin et al, 2010).…”

“…Morever, addition of different carban sources increased the paclitaxel production (Ketchum and Gibson, 1996;Ketchum et al, 2003). Use of the biotic and abiotic elicitors to improve taxol production has been studied (Strobel et al, 1992;Ciddi et al, 1995;Yukimune et al, 1996;Jha et al 1998). Ellis et al, (1996) established cultures using nodal segment in seven Taxus cultivars and screened the taxol production.…”

“…The major advantages of cell cultures includes (i) synthesis of bioactive secondary metabolites independently from climatic and soil conditions; (ii) negative biological influences that affect secondary metabolites production in the nature are eliminated (microorganisms and insects) (iii) to select cultivars with higher production of secondary metabolites; (iv) with automatization of cell growth control and metabolic processes regulation, cost price can decrease and production increase (Jha et al, 1998;Abdin et al 2003;Junaid et al, 2009;Junaid et al, 2010). The objectives of many industries are to develop plant cell culture techniques to the stage where they yield secondary products, more cheaply than extracting either the whole plant grown under natural conditions or synthesizing the product.…”